Research On The Mode Of In Vivo Synthesis Of Gold Nanoclusters Based On The Self-made Micro Culture System

For a long time,cancer has become the most serious disease that endangers the health of our residents.Early diagnosis and targeted therapy of cancer have become the focus of researchers.Identification and analysis of tumor cells is an important research tool in early diagnosis of cancer.Biological imaging technology is a research field with broad application prospects in biomedical engineering.As a cross discipline,it combines molecular biology with advanced imaging techniques and uses specific molecular targeting probes to achieve tracking imaging,so that molecules,genes and cells are studied qualitatively and quantitatively.Optical bio-imaging is widely used in biomedical research for its advantages of high spatial and temporal resolution,fast imaging speed,good sensitivity,low cost and no radioactive hazards.In recent years,the imaging technology based on biosynthesis in vivo has attracted wide attention from researchers for it is noninvasively and simply.However,the existing biological imaging technology often separates the biological culture from the imaging,which limits the advantages of the biosynthesis in situ,and cannot guarantee the vitality of the organism in the imaging.Therefore,this paper has designed a mirco culture system,which can be placed on a microscope platform.By controlling the temperature,humidity and CO₂ concentration of the miniature incubator,the cell culture was realized.By combining the micro culture system with the confocal fluorescence microscope,the location and intensity of the fluorescence of the tumor cells were studied when the gold nanoparticles were synthesized in vivo.The main contents of this paper are as follows:A mirco culture system,which can be placed on the inverted microscope,was designed and produced.The temperature,humidity and CO₂ concentration can be accurately controlled by sensing and PID control technology,maintaining a stable and efficient cell culture environment.Through serial communication with the computer,we can record the temperature,humidity and CO₂ concentration information of self-product incubator.The experimental results show that the device can control the temperature,humidity and CO₂ concentration.The cell viability was tested by MTT assay,compared with the common CO₂ culture box,the self-product culture system and ordinary indoor conditions.We found that the cell vitality in the self-product culture system reached more than 95%of the control group.At the same time,the cell viability of ordinary indoor conditions was less than 50%.Therefore,we believe this self-product culture system can provide a stable and efficient extracellular environment for cell incubation.We applied the micro culture system to the cells imaging,using in vivo biosynthesis of gold nanoclusters to establish a self-imaging model.The experimental results showed that the cell fluorescence was mainly concentrated in the endoplasmic reticulum area of the tumor cells,and closely related to the activity and energy state of the tumor cells.The luminance was high and full of the whole cytoplasm,when metabolic activity of cells was strong.Fluorescence in cells was markedly different at different stages of cell cycle.When added the Chlorchloric acid into culture medium in the G0 phase,the fluorescent substances in the cells mainly shrink in a region near the nucleus.When added the Chlorchloric acid into culture medium in the G2 phase the fluorescent nanoclusters were brighter than other phases,mainly because the cell metabolism was more energetic,and more substances were absorbed into the cells.In contrast,normal cells have relatively weak fluorescence due to their long cell cycle and poor metabolic activity.